Engineered protein kinases which can utilize modified nucleotide triphosphate substrates

Abstract

Engineered protein kinases which can utilize modified nucleotide triphosphate substrates that are not as readily utilized by the wild-type forms of those enzymes, and methods of making and using them are disclosed. Modified nucleotide triphosphate substrates and methods of making and using them are disclosed. Methods are disclosed for using such engineered kinases and such modified substrates to identify which protein substrates the kinases act upon, to measure the extent of such action, and to determine if test compounds can modulate such action. Engineered forms of multi-substrate enzymes which covalently attach part or all of at least one (donor) substrate to at least one other (recipient) substrate, which engineered forms will accept modified substrates that are not as readily utilized by the wild-type forms of those enzymes are disclosed. Methods for making and using such engineered enzymes are disclosed. Modified substrates and methods of making and using them are disclosed. Methods are disclosed for using such engineered enzymes and such modified substrates to identify the recipient substrates the enzymes act upon, to measure the extent of such action, and to measure whether test compounds modulate such action.

1. A nucleotide sequence which encodes a mutant methyl transferase enzyme which accepts an S-adenosylmethionine analog as a methyl donor substrate, whereby catalytic activity of said enzyme results in the transfer of a methyl group of said substrate to at least one other substrate of said enzyme. 
     
     
       2. A method for producing a mutant methyl transferase enzyme which accepts an S-adenosylmethionine analog as a methyl donor substrate, comprising expressing a the nucleotide sequence of  claim 1  whereby said mutant methyl transferase enzyme is produced.